New indication of paroxetine pharmaceutical composition for treating cancer

ABSTRACT

A method for treating a cancer includes administering to a subject in need thereof a pharmaceutical composition containing a therapeutically effective amount of Paroxetine or a pharmaceutical acceptable salt thereof. The cancer is selected from the group consisting of pleural-related cancer, abdominal-related cancer, endocrine-related cancer, gastrointestinal tract-related cancer, osteosarcoma, skin cancer, and blood cancer. The pleural-related cancer is lung cancer. The abdominal-related cancer is selected from bladder cancer, cervical cancer, and kidney cancer. The endocrine-related cancer is selected from prostate cancer, breast cancer, and ovarian cancer. The gastrointestinal tract-related cancer is selected from gastric cancer, hepatic cancer, colorectal cancer, pancreatic cancer, and tongue cancer.

CROSS-REFERENCE TO RELATED APPLICATIONS

This is a National Phase Application filed under 35 U.S.C. 371 as anational stage of PCT/CN2015/092780 filed Oct. 23, 2015, an applicationclaiming the benefit under 35 USC 119(e) to the following U.S.Provisional Applications No. 62/068,298 filed Oct. 24, 2014, the contentof each of which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention related to a new indication of Paroxetinepharmaceutical composition, especially related to inhibition effect ofParoxetine pharmaceutical composition on a variety of cancer cells.

BACKGROUND OF THE INVENTION

Cancer is the most popular disease cause of death in the world. Thecancer patients are gradually increase yearly, therefore the treatmentmethod of the cancer has become an important issue. The medicaltreatments of cancer can be classified as surgical treatment, radiationtherapy, chemotherapy and target therapy. Generally, the cancer drug,whether chemotherapy drug or target therapy drug, is to inhibit cancercells duplication and split to prevent the tumor growth and metastasis.

Recently, the drug design for cancer is mainly focused on development ofhigh-specific drug molecules or target-based antibody. Averagely, onlyabout five of 10,000 new drugs can successfully enter the phase I ofclinical trials.

Otherwise, the manufacturing of the drug is also a big problem. When thedrug starting the clinical trials, there are lots of problems need toovercome, such as drug safety, patient selection, trial dose and otherissues. Even the drug has approved by the FDA and sales on the market,there still possibly face the situation of the poor drug response inpatients. Furthermore, if the cancer patients happen the drugresistance, that would reduce the effectiveness of the drugs and resultin the medical treatment failure. Therefore, the new drug development isvery difficult.

Paroxetine, also known by the trade names Paxil and Seroxat, is anantidepressant of the selective serotonin reuptake inhibitor (SSRI)class. It is used to treat major depressive disorder,obsessive-compulsive disorder, social anxiety disorder, panic disorder,posttraumatic stress disorder, generalized anxiety disorder andpremenstrual dysphoric disorder. Paroxetine is approved by FDA andaccumulated a huge data of drug use and drug mechanism research.

Due to the differences of the clinical use, there is no research presentthat the paroxetine has any potential to inhibit cancer cell.

SUMMARY OF THE INVENTION

In order to solve the above problems, the present invention provide thedevelopment of new cancer clinical indications of Paroxetine.

Accordingly, the present invention provides a new indication ofParoxetine. The experimental results showed that the Paroxetine had notoxicity or had little toxicity to normal cells in the presentinvention.

The present invention provides a pharmaceutical composition ofParoxetine for treating cancer. The pharmaceutical composition iscomposed of effective dose of Paroxetine and a pharmaceutical acceptablesalt.

In one embodiment of the present invention, the cancer is selected frompleural-related cancer, abdominal-related cancer, endocrine-relatedcancer, gastrointestinal tract-related cancer.

In one embodiment of the present invention, the cancer is selected fromosteosarcoma, skin cancer and blood cancer.

In one embodiment of the present invention, the pleural-related canceris lung cancer.

In one embodiment of the present invention, the abdominal-related canceris selected from bladder cancer, cervical cancer and kidney cancer.

In one embodiment of the present invention, the endocrine-related canceris selected from prostate cancer, breast cancer, and ovarian cancer.

In one embodiment of the present invention, the gastrointestinaltract-related cancer is selected from gastric cancer, hepatic cancer,colorectal cancer, pancreatic cancer, and tongue cancer.

In one embodiment of the present invention, the effective dose ofParoxetine is from 20 mg/kg/day to 500 mg/kg/day.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the results of the inhibitory effect of the differentcancer cells by Paroxetine.

FIG. 2 shows the results of the inhibitory effect of gastric tumorvolume by Paroxetine.

FIG. 3 shows the inhibitory effect of gastric tumor growth viaadministered high-dose and low-dose of Paroxetine.

DETAILED DESCRIPTION OF THE INVENTION

Cell Culture

Subculture the different types of cancer cells. The cancer cellsincludes lung cancer, gastric cancer, hepatic cancer, colon cancer, skincancer, cervical cancer, prostate cancer, bladder cancer, breast cancer,leukemia, pancreatic cancer, ovarian cancer, tongue cancer,osteosarcoma, and renal cancer. The normal cells used in the controlgroup included kidney cells (HEK293), canine fibroblast cell line HFW,and human bronchial epithelial cell line BEAS-2B. (as shown in Table 1).

Cancer cell lines were cultured in different culture medium according todifferent characteristics (as shown in Table 1). The cell numbers werecounted and reseed as 2×10⁶ in culture plate/flask. Then, the culturemedium were added to a volume of 10 ml, and the cells were cultured for2-3 days. Then, the cells were suspended for loading into 96-wellplates. The number of cells was 3000 and the volume of the culturemedium was 100 μl each well.

TABLE 1 Cancer cell lines and culture medium Culture No Cancer typeCancer cell type medium 1 lung cancer H1650 (lung adenocarcinoma)RPMI-1640 A549 (lung adenocarcinoma) DMEM 2 gastric cancer AGS (GastricAdenocarcinoma) RPMI-1640 MKN-45 (Gastric RPMI-1640 Adenocarcinoma) 3hepatic cancer HepG2 (hepatocellular DMEM carcinoma) DMEM Hep3B(hepatocellular carcinoma) 4 colon cancer HCT116(p53+) (colorectal DMEMcarcinoma) DMEM LoVo(Colorectal Adenocarcinoma) 5 skin cancer A375(amelanotic melanoma) DMEM BCC (basal cell carcinoma) DMEM 6 cervicalcancer HeLa (CervixAdenocarcinoma) DMEM C-33A (Cervical carcinoma) MEMBCRC60554 7 prostate cancer PC3 (p53−)(Prostate DMEM adenocarcinoma)RPMI-1640 LNCaP clone FGC (LNCap.FGC) 8 bladder cancer 8301 (urinarybladder RPMI-1640 carcinoma) RPMI-1640 T24 9 breast cancer MCF7 (MammaryGland, DMEM Adenocarcinoma) DMEM MDA-MB-231 (Mammary Gland,Adenocarcinoma) 10 pancreatic cancer BxPC-3 RPMI-1640 AsPC-1 RPMI-164011 ovarian cancer NIH: OVCAR-3 RPMI-1640 TOV-21G RPMI-1640 12 tonguecancer SAS (Tongue squamouscell DMEM carcinoma) 13 osteosarcoma U-2OSDMEM 14 renal cancer 786-O (Renal adenocarcinoma) RPMI-1640 BCRC 6024315 normal cell kidney HEK293 (Kidney), MDCK, DMEM pulmonary VERORPMI-1640 epithelial cell line BEAS-2B (Lung Epithelial)

Cell Viability Analysis

Removing the original culture medium from 96-well plate. Then add 100 μlof commercially drug at a concentration of 10 μM per well. After 72hours, add the diluted WST-1 reagent to the well with 100 μl/well, andthe diluted WST-1 reagent was acquired from the dilution of 9:1 mediumand WST-1 stock reagent. Finally, the total volume of each well was 200μl/well. Culture the 96-well plate at 37° C. for 30 to 90 minutes.Detecting and calculate the survival rate of each cancer cells with anELISA reader at OD450 nm. The lower viability of cancer cells representsbetter inhibition effect via the Paroxetine drug. Otherwise, the higherviability of cancer cells represents worse inhibition effect via theParoxetine drug.

The Effect of Paroxetine on Different Cancer Cell Lines

The Inhibition Effect of Paroxetine on Pleural-Related Cancer Cells

This inhibition test of Paroxetine on pleural-related cancer cells wereusing two lung cancer cell lines A549 and H1650. The inhibitory tests ofParoxetine were performed 4 times for each cell lines and then theaverage value of the inhibitory tests was calculated. The results wereshown in Table 2.

TABLE 2 The inhibition effect of Paroxetine on pleural-related cancercell lines 0524-10 0526-10 0529-10 0531-10 min min min min Average A54984.19 127.41 54.01 104.12 92.43 1-10 2-20 3-20 4-20 min min min minAverage H1650 66.4 31.6 58.2 71.3 56.9

The Inhibition Effect of Paroxetine on Abdominal-Related Cancer CellLines

This inhibition test of Paroxetine on abdominal-related cancer cellswere using bladder cancer cell lines TSGH and T24 (Table 3), cervicalcancer cell lines HeLa and C-33A (Table 4), renal cancer cell line 786-O(Table 5).The inhibitory tests of Paroxetine were performed 4 times foreach cell lines and then the average value of the inhibitory tests wascalculated. The results were shown in Table 3, Table 4, and Table 5.

TABLE 3 The inhibition effect of Paroxetine on bladder cancer cell lines0510-10 0512-10 0515-10 0517-10 min min min min Average TSGH 37.09 37.3936.75 42.18 38.4 1-30 2-20 3-20 4-20 min min min min Average T24 111.785.2 101.5 101.5 99.9

TABLE 4 The inhibition effect of Paroxetine on cervical cancer celllines 0524-10 min 0526-10 min 0529-10 min 0531-10 min Average HeLa 52.6178.32 50.99 73.36 63.82 C- 33.2 32.1 31.7 35.8 33.2 33A

TABLE 5 The inhibition effect of Paroxetine on renal cancer cell line0524-10 min 0526-10 min 0529-10 min 0531-10 min Average 786- 35.2 24.014.6 22.3 24.0 O

The Inhibition Effect of Paroxetine on Endocrine-Related Cancer CellLines

This inhibition test of Paroxetine on endocrine-related cancer cellswere using prostate cancer cell lines PC-3 and LNCap (Table 6), breastcancer cell lines MCF7 and MDA-MB-231 (Table 7),and ovarian cancer celllinesNIH-OVCAR-3 and TOV-21G(Table 8). The inhibitory tests ofParoxetine were performed 4 times for each cell lines and then theaverage value of the inhibitory tests was calculated. The results wereshown in Table 6, Table 7, and Table 8.

TABLE 6 The inhibition effect of Paroxetine on prostate cancer celllines PC-3-0524- PC-3-0526- PC-3-0529- PC-3-0531- Aver- 10 min 10 min 10min 10 min age PC-3 33.52 61.61 39.26 41.84 44.06 LNCap-1- LNCap-2-LNCap-3- LNCap-4- 10 min 20 min 20 min 20 min Average LNCap 65.9 61.780.9 78.3 71.7

TABLE 7 The inhibition effect of Paroxetine on breast cancer cell lines0619- 0612-10 min 0614-10 min 0616-10 min 10 min Average MCF7 47.7149.97 55.41 51.60 51.17 MDA- 47.94 35.88 42.04 41.18 41.76 MB-231

TABLE 8 The inhibition effect of Paroxetine on ovarian cancer cell lines7-3-30 min 7-4-30 min 7-7-30 min Average 7-4- 30 min NIH- 80.2 81.9 83.997.8 86.0 OVCAR-3 4-30 min TOV-21G 93.5 79.7 89.2 86.1 87.1

The Inhibition Effect of Paroxetine on Gastrointestinal Tract-RelatedCancer Cell Lines

This inhibition test of Paroxetine on gastrointestinal tract-relatedcancer cells were using gastric cancer cell lines AGS and MKN-45 (Table9), hepatic cancer cell lines HepG2 and Hep3B (Table 10), colorectalcancer cell lines HCT116-wt and LoVo (Table 11), pancreatic cancer celllines AsPC-1 and BxPC-3 (Table 12), and tongue cancer cell line SAS(Table 13).The inhibitory tests of Paroxetine were performed 4 times foreach cell lines and then the average value of the inhibitory tests wascalculated. The results were shown in Table 9, Table 10, Table 11, Table12and Table 13.

TABLE 9 The inhibition effect of Paroxetine on gastric cancer cell lines0517- 0510-10 min 0512-10 min 0515-10 min 10 min Average AGS 12.02 11.1012.65 12.85 12.2 MKN- 36.20 52.00 37.76 36.02 40.5 45

TABLE 10 The inhibition effect of Paroxetine on hepatic cancer celllines 0531- 0524-20 min 0526-20 min 0529-20 min 20 min Average HepG266.44 58.28 50.07 41.73 54.13 0619- 0612-20 min 0614-20 min 0616-20 min20 min Average Hep3B 69.85 67.38 66.16 60.90 66.07

TABLE 11 The inhibition effect of Paroxetine on colorectal cancer celllines 0609- 0602-30 min 0605-10 min 0607-10 min 10 min Average HCT116-23.78 30.62 29.66 29.47 28.38 wt 0623- 0616-10 min 0619-10 min 0621-10min 10 min Average LoVo 54.11 51.54 77.28 48.93 57.97

TABLE 12 The inhibition effect of Paroxetine on pancreatic cancer celllines 1-4- 1-7-3-30 min 1-7-4-30 min 1-7-7-30 min 30 min Average AsPC-178.8 87.3 69.3 72.5 77.0 3-4- 3-7-3-30 min 3-7-4-30 min 3-7-7-30 min 30min Average BxPC-3 59.7 93.7 70.6 87.0 77.7

TABLE 13 The inhibition effect of Paroxetine on tongue cancer cell line6-26-10 min 6-28-10 min 6-30-10 min 7-3-10 min Average SAS 48.20 26.7457.97 73.09 51.50

The Inhibition Effect of Paroxetine on Other Cancer Cell Lines

This inhibition test of Paroxetine on other cancer cells were usingosteosarcoma cell line U2OS (Table 14), skin cancer cell lines A375 andBCC (Table 15). The inhibitory tests of Paroxetine were performed 4times for each cell lines and then the average value of the inhibitorytests was calculated. The results were shown in Table 14 and Table 15.

TABLE 14 The inhibition effect of Paroxetine on osteosarcoma cancer cellline 7-3- 6-26-10 min 6-28-10 min 6-30-10 min 10 min Average U2OS 40.6130.75 30.29 31.10 33.19

TABLE 15 The inhibition effect of Paroxetine on skin cancer cell lines0609- 0602-30 min 0605-10 min 0607-10 min 10 min Average A375 46.7544.40 35.38 48.77 43.83 BCC 42.13 52.62 41.11 97.05 58.23

The Experiment Design on Control Group

The Inhibition Effect of Paroxetine on Normal Cells

This inhibition test of Paroxetine on normal cells were using normalkidney cell line HEK293 (Table 16) and normal pulmonary epithelial celllines BEAS-2B (Table 17). The inhibitory tests of Paroxetine wereperformed 4 times for each cell lines and then the average value of theinhibitory tests was calculated. The results were shown in Table 16 andTable 17.

TABLE 16 The inhibition effect of Paroxetine on normal kidney cell line0609- 0602-30 min 0605-30 min 0607-30 min 30 min Average HEK293 64.9176.80 76.05 67.85 71.40

TABLE 17 The inhibition effect of Paroxetine on normal pulmonaryepithelial cell line 0517- 0510-10 min 0512-10 min 0515-10 min 10 minAverage BEAS-2B 100.97 102.64 96.40 94.56 98.64

This inhibition test results of Paroxetine on all kinds of cells wereshown in Table 18. It is clear that Paroxetine has a significantinhibitory effect on several cancer cell lines. As a result in theexperiments of the present invention, Paroxetine has a significantinhibitory effect on various cancer cells. (FIG. 1)

TABLE 18 Summary of the Effect on different cancer cell lines byParoxetine cancer cells Inhibitory effect lung cancer 74.64462204bladder cancer 69.1 cervical cancer 63.82 prostate cancer 44.06 breastcancer 46.47 ovarian cancer 86.5 gastriccancer 26.3 hepatic cancer 60.10colorectal cancer 43.17 pancreatic cancer 77.4 tongue cancer 51.50osteosarcoma 33.19 skin cancer 51.03 renal cancer 72.5 kidney cell 71.40pulmonary epithelial cell line 98.64

Animal Model Test of Gastric Cancer with Dose 100 mg/kg/day and 200mg/kg/day

In this invention, the female mice were(BALB/cAnN.Cg-Foxn1^(nu)/CrlNarl) purchased from National LaboratoryAnimal Center (Taiwan). The weight of the mice were 21±1 g. These micewere subcutaneously injected with gastric cancer cells (AGS) and thenput these mice into different cage sat random. The drug test experimentwas divided into three groups, include “control group”, “low dose group(100 mg/kg/day)”, and “high dose group (200 mg/kg/day)”. These mice werethen injected test drug intraperitoneally once daily until the tumorsize reached 100 mm³. The tumor sizes and body weight were measuredtwice a week. The tumor sizes were measured and calculated by formula:(L×W²)/2. L represents the tumor longest length. W represents the tumorshortest diameter. The experiment results were shown in Table 19.

TABLE 19 The inhibitory effect of tumor volume via administeredParoxetine control group Tumor low concentration (100 mg/kg/day) longestvolume longest weight length width volume growth weight length widthvolume (g) mm mm mm3 mm3 (g) mm mm mm3 First measurement A 18.5 7 7171.5 171.5 19 7 5 87.5 B 22 8 6 144 144 19 8 6 144 C 20.5 9 8 288 28818 6 6 108 average 201.1667 201.1667 113.1667 Second measurement A 22 76 126 −45.5 19 6 5 75 B 20 8 7 196 52 21 7 6 126 C 20 9 7 220.5 −67.5 207 6 126 average 180.8333 −20.3333 109 Third measurement A 23 9 6 162 3620.5 7 6 126 B 20 10 8 320 124 23.5 7 4 56 C 21 11 7 269.5 49 19 7 587.5 average 250.5 69.667 89.833 Fourth measurement A 23 11 7 269.5107.5 22 6 6 108 B 22 10 6 180 −140 22 4 4 32 C 23 11 8 352 82.5 20 6 575 average 267.1667 16.667 71.667 Fifth measurement A 22 12 8 384 114.522 6 6 108 B 22 11 8 352 172 20 4 5 50 C 23 12 9 486 134 20 4 4 32average 407.33 140.167 63.333 high concentration (200 mg/kg/day) Tumorlongest volume low concentration (100 mg/kg/day) weight length widthvolume growth Tumor volume growth mm3 (g) mm mm mm3 mm3 Firstmeasurement A 87.5 19 6 4 48 48 B 144 18.5 8 5 100 100 C 108 20 7 5 87.587.5 average 113.1667 78.5 78.5 Second measurement A −12.5 20 6 4 48 0 B−18 23 5 3 22.5 −77.5 C 18 20 7 5 87.5 0 average −4.16667 52.66667−25.8333 Third measurement A 51 19 5 4 40 −8 B −70 18.5 0 0 0 −22.5 C−38.5 18.5 0 0 0 −87.5 average −19.167 13.333 −39.33 Fourth measurementA −18 20 0 0 0 −40 B −24 21 0 0 0 0 C −12.5 20 0 0 0 0 average −18.167 0−13.33 Fifth measurement A 0 21 0 0 0 0 B 18 20 0 0 0 0 C −43 20 0 0 0 0average −8.333 0 0

According to the results in FIG. 2, both low dose and high dose ofParoxetine had significant inhibition effect on tumor cells, and theweight of mice did not show a significant decrease during theexperiment. These results indicate that both high and low doses ofParoxetine could keep the tested mice in healthy status during thetreatment without death.

According to the results in FIG. 3, both low dose and high dose ofParoxetine had effectively slow down the tumor volume growth, and canalso reduce the tumor volume. Especially, high doses of Paroxetine hadbetter effect to inhibit tumor growth.

Although the present invention has been described in terms of specificexemplary embodiments and examples, it will be appreciated that theembodiments disclosed herein are for illustrative purposes only andvarious modifications and alterations might be made by those skilled inthe art without departing from the spirit and scope of the invention asset forth in the following claims.

1. A method for treating a cancer comprising: administering to a subjectin need thereof a pharmaceutical composition comprising atherapeutically effective amount of Paroxetine or a pharmaceuticalacceptable salt thereof.
 2. The method of claim 1, wherein the cancer isselected from the group consisting of pleural-related cancer,abdominal-related cancer, endocrine-related cancer, and gastrointestinaltract-related cancer.
 3. The method of claim 1, wherein the cancer isselected from the group consisting of osteosarcoma, skin cancer, andblood cancer.
 4. The method of claim 2, wherein the pleural-relatedcancer is lung cancer.
 5. The method of claim 2, wherein theabdominal-related cancer is selected from the group consisting ofbladder cancer, cervical cancer, and kidney cancer.
 6. The method ofclaim 2, wherein the endocrine-related cancer is selected from the groupconsisting of prostate cancer, breast cancer, and ovarian cancer.
 7. Themethod of claim 2, wherein the gastrointestinal tract-related cancer isselected from the group consisting of gastric cancer, hepatic cancer,colorectal cancer, pancreatic cancer, and tongue cancer.
 8. The methodof claim 1, wherein the effective amount of Paroxetine is from 20mg/kg/day to 500 mg/kg/day.